title: Cloning and characterization of a small family 19 chitinase from moss (Bryum coronatum).
abstract: Chitinase-A (BcChi-A) was purified from a moss, Bryum coronatum, by several steps of column chromatography. The purified BcChi-A was found to be a molecular mass of 25 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an isoelectric point of 3.5. A cDNA encoding BcChi-A was cloned by rapid amplification of cDNA ends and polymerase chain reaction. It consisted of 1012 nucleotides and encoded an open reading frame of 228 amino acid residues. The predicted mature BcChi-A consists of 205 amino acid residues and has a molecular weight of 22,654. Sequence analysis indicated that BcChi-A is glycoside hydrolase family-19 (GH19) chitinase lacking loops I, II, IV and V, and a C-terminal loop, which are present in the catalytic domain of plant class I and II chitinases. BcChi-A is a compact chitinase that has the fewest loop regions of the GH19 chitinases. Enzymatic experiments using chitooligosaccharides showed that BcChi-A has higher activity toward shorter substrates than class II enzymes. This characteristic is likely due to the loss of the loop regions that are located at the end of the substrate-binding cleft and would be involved in substrate binding of class II enzymes. This is the first report of a chitinase from mosses, nonvascular plants.
authors.affiliation: Department of Bioscience and Biotechnology, Faculty of Agriculture, University of the Ryukyus, Okinawa, Japan. tokey@agr.u-ryukyu.ac.jp
authors.forename: Toki
authors.initials: T
authors.lastname: Taira
authors.forename: Yoko
authors.initials: Y
authors.lastname: Mahoe
authors.forename: Noriko
authors.initials: N
authors.lastname: Kawamoto
authors.forename: Shoko
authors.initials: S
authors.lastname: Onaga
authors.forename: Hironori
authors.initials: H
authors.lastname: Iwasaki
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ohnuma
authors.forename: Tamo
authors.initials: T
authors.lastname: Fukamizo
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D019068:Bryopsida / Q000201:enzymology*; D020134:Catalytic Domain; D002688:Chitinases / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D003001:Cloning, Molecular; D057075:Enzyme Assays; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D014242:Trichoderma / Q000187:drug effects
chemical_list: D000935:Antifungal Agents; D002688:Chitinases